ALLERGY ANALYSIS: Alternative Methods

By Stephen G. Perry Ph.D.

I had an opportunity to establish a Lowry quantitative histochemical laboratory at the Missouri Medical School Missouri Institute of Psychiatry (MIP) in Saint Louis Missouri as an Assistant Professor. While Director of the Missouri Division of Mental Health Dr. George A. Ulett, M.D., Ph.D. established MIP to provide research into the causes of mental problems. As the neurochemistry lab director Dr. Ulett asked me to direct his allergy research program we undertook some really challenging research problems. His wife, Dr Pearl Ulett, Child Psychiatrist, was conducting food allergy and diet management studies that reversed the symptoms that got youth diagnosed with mental problems and a side benefit of loss of acne and weight gain. I coordinated a family study beginning with mine and submitted myself to a food allergy computer analyzed EEG study with spectacular findings. I also recommended a comprehensive research plan to include food allergies and various diagnosed patients versus control groups. 

• Language: English
• Publisher: Outskirts Press
• Release date: May 2, 2023
• ISBN: 9781977264572

Stephen G. Perry Ph.D.

My first job was an Assistant Professor and Director of the Neurochemistry Laboratory. This book tells the story of our findings. With positive foods WBCs go up while negative foods match the zero-time controls. When white cells go up so does the immune system including plasma white cells that recognize proteins and generate monoclonal antibodies some of which can be autosomal. When Dr Ulett was no longer Missouri Mental Health Director this research went away.    

Book preview

ALLERGY ANALYSIS: Alternative Methods

All Rights Reserved.

Copyright © 2023 Stephen G. Perry Ph.D.

v10.1

The opinions expressed in this manuscript are solely the opinions of the author and do not represent the opinions or thoughts of the publisher. The author has represented and warranted full ownership and/or legal right to publish all the materials in this book.

This book may not be reproduced, transmitted, or stored in whole or in part by any means, including graphic, electronic, or mechanical without the express written consent of the publisher except in the case of brief quotations embodied in critical articles and reviews.

Outskirts Press, Inc.

http://www.outskirtspress.com

ISBN: 978-1-9772-6457-2

Cover Photo © 2023 Stephen G. Perry Ph.D. All rights reserved - used with permission.

Outskirts Press and the OP logo are trademarks belonging to Outskirts Press, Inc.

PRINTED IN THE UNITED STATES OF AMERICA

Contents

Preface

CHAPTER 1: Historical

CHAPTER 2: First Analysis

CHAPTER 3: Single Sensitizing Foods

CHAPTER 4: Cerebral Allergy

CHAPTER 5: Familial Relationships

CHAPTER 6: Population Analysis

CHAPTER 7: Summary and Conclusions

Preface

It has been estimated that a significant percentage of the population exhibit clinical symptoms due directly to food sensitivities with the possibility that many more suffer from other allergies or ill-defined medical problems that may at least be aggravated by their diets. Then, as now, a number of clinical tests aimed at aiding the clinician in the diagnosis of allergy problems are not definitive and do not provide for effective screening and isolation of unfavorable symptomatology. In some cases, sophisticated and highly advanced procedures have not benefited from systematic application and follow-up study to appreciate fully the advances made. In the mid 1970’s we reported that the in vitro Cytotoxic Test correlated with in vivo food challenges and was an excellent screening technique to identify allergens that affect the skin, and potentially other organ systems such as the cardiovascular, gastrointestinal, respiratory, and central nervous. In this book ALLERGY ANALYSIS ALTERNATIVE METHODS, we offer a follow-up study for the results of systematic Cytotoxic Testing. Data is presented that correlates Cytotoxic Test results with clinical findings, familial distributions, and population studies. This book is written in detail to clarify and further substantiate our earlier reports (Ulett, GA, and SG Perry 1974; 1975) and ongoing studies. The detail should also permit other scholars to analyze the data according to their fields of expertise. I also recommend this book to students and practicing clinicians as a guide for interpreting single diagnostic tests and to plan testing and treatment regimens to better serve the patient’s well-being and preventative health programs.

I dedicate this book to George A. Ulett, Ph.D. M.D. whose vision created the Missouri Institute of Psychiatry (MIP) and provided inspired leadership to MIP and its many programs, some of which achieved international recognition.

Sharon Authenreith-Netherly deserves considerable recognition for her skills as a technician and her dedication to the patients and volunteers that participated in Dr Ulett’s programs.

Of particular recognition are the many that participated in the program and volunteered of themselves and their families then spread the word in the community for others to come and seek treatment from complaints of discomfort that they suffered and yet had not found relief. If you found them today, they are still probably talking about the commonsense benefit that they and family members received.

It has taken all too long to tell this story!

CHAPTER 1

Historical

BLACK, M.D. (1956), an El Paso, Texas Pediatrician that described the in vitro (in glass) effects of allergens A New Diagnostic Method in Allergic Disease was a benchmark publication by Arthur P. on white blood cells of allergic children. The defining clinical event that led Dr. Black to investigate and conceptualize his diagnostic methodology came in 1928 when a blood transfusion he administered led to acute agranulocytosis and subsequent death of a child. Black reported that prior to transfusion the child had marked leukocytosis (rising WBC) with a high percentage of polymorph nuclear cells (segmented neutrophils). To understand, the adverse reaction Dr. Black used donor and recipient bloods remaining from cross matching to prepare warm stage slides with supravital staining. Under the microscope, he observed the destruction of the recipient’s polymorphonuclear leukocytes in 30 - 40 minutes while control cell preparations remained viable and active. Dr. Black found no explanation in 1928; however, in 1956 he speculated that the phenomenon might have been an allergic one...in which the donor’s plasma contains an allergen, the child’s plasma, a homologous antibody, and the leukocytes act as the shock organ". His search of the literature for evidence describing the in vitro behavior of leukocytes in the presence of both allergen and reaginic plasma from a sensitized individual yielded only papers reporting pieces of that puzzle. He noted that Hektoen (1906) reported in a study of bacterial phagocytosis that the variable factor is the serum and not the leukocytes. Hektoen also demonstrated that antibody can be eluted from leukocytes in salt solutions. Studying bacterial hypersensitivity Blatt and Nantz (1955) used washed cells relatively void of plasma and antibody in experiments employing leukocytes and plasma with no leukocytotoxic effect or leukocytes and antigen with no leukocytotoxic effect, but not leukocytes, plasma, and allergen together in a single preparation. Prausnitz and Kustner (1921) demonstrated this three-way requirement in dramatic fashion by transferring skin sensitizing antibodies from an allergic person to the skin of a nonallergic person, then using these inoculated leukocyte containing sites for skin tests with specific antigens. Described as a highly reliable diagnostic test the P-K test is impractical for many reasons and therefore this passive transfer testing procedure applies only in exceptional circumstances. These observations laid the foundation for Black (1956) to develop a new method, referred to as the Cytotoxic Test, with ten supporting clinical case studies.

The Cytotoxic Test: Black (1956) provided a list of recommended materials for Cytotoxic Testing. The leukocytes were concentrated and separated from erythrocytes with the use of acid-citrate-dextrose anticoagulant and centrifugation. The original procedure recommended only heparinized hematocrit tubes with anticoagulant taken up by capillary action to 6-8 mm followed by a finger or heel stick sample to within a cm of the end of the tube. To ensure the admixture of blood and anticoagulant a rubber bulb attached to the tube and its contents expelled into a hollow ground slide and reaspirated several times. The crit tube(s) were refilled, sealed, and centrifuged 15 - 30 minutes at 1200 rpm. Scratch each tube at the bottom of the buffy coat and discard the erythrocytes. The contents of the tube(s) are admixed in a hollow ground slide then taken up in plain capillary tube. Two tubes provided enough plasma-leukocyte cell suspension for six tests. Allergens were prepared by placing a drop (~ 0.1 ml) of distilled water on a slide and mixing an amount of dried allergen (Holister-Stier Labs, same as used for skin testing) transferred on the narrow end of a toothpick then allowed to dry in an area less than a circular cover slip. If neutral red supravital staining (25 mgs/100 mls ETOH) is going to be used place a drop in the center of the cover slip and allowed to dry. At the time of testing select the appropriate slide(s), add a drop (approx. 0.1 ml) of plasma-leukocyte cell suspension on the allergen matrix of the slide then cover slipped and sealed with a Vaseline-mineral oil sealant. Incubate at 37 C observing under a warm stage microscope at 30 min intervals. Carry controls, which are viable and stable under these conditions for hours through the same procedures. The Cytotoxic effects manifest as progressive changes seen in the polymorphonuclear leukocytes: as a loss in ameboid activity, a rounding of cell contour, a decrease and cessation of cytoplasmic movement, and increased staining. Erythrocytes and other cellular elements of the blood are generally not affected. Black developed this procedure as a diagnostic tool to support his practice. His case reports (Black, 1956) indicated modest testimonial success.

The Bryans (1960) employed the Cytotoxic food allergy test, varying only slightly from the method of Black (1956), as one of many diagnostic tools in their global assessment of patients presenting themselves to their otolaryngology practice associated with Barnes Hospital/Washington University Medical School, St. Louis, Missouri. They improved the techniques for isolating viable leukocytes in such quantities to support large-scale screening of patients for a broader spectrum of potential allergens. They standardized the buffy coat/plasma mixture to ensure 15 - 40 leukocytes per microscopic (60X apochromatic high dry objective) field. Since cells settle rapidly precautions are required in their procedures to gain consistency of handling and distributing the buffy coat cell suspensions. The standardized antigen preparations 0.1% (1.0mg/1.0 ml) solutions are matched with the dried matrix on the slide being slightly larger than the cover slip for more uniformity with batches of test slides made in advance. Clean Silicad glassware, slides and cover slips were a prerequisite for performing the test especially improving the controls where unfavorable protein binding to glass may occur. Performing the test at room temperature (80 - 85 F) at determined intervals proved acceptable and permitted more uniformity with batch screening. The Bryan’s (1960) evaluated the degree of cytotoxic reaction as negative, or positive (slight, moderate, or marked). The Bryan’s described the Cytotoxic reactions of the neutrophils as presenting the following changes: 1) loss of amoeboid movement and cessation of streaming of cytoplasmic granules; 2) rounding of the cell contour with or without spine-like projections; 3) vacuolization and spreading of the cytoplasm where the cell may become thin and flat; and 4) fragmentation of the cell with rupture of the membranes and dispersion of the granules. A negative cytotoxic reaction shows no changes as described above and are consistent with the no changes seen in the controls carried with each experiment (i.e., nonreactive foods, or distilled water). A slight positive shows half of the leukocytes losing their amoeboid activity and displaying a rounded contour upon scanning 10 - 15 fields. A moderate positive cytotoxic reaction shows all of the leukocytes lose their amoeboid activity and display a rounded contour with a few cells disintegrating. A marked positive cytotoxic reaction shows the majority of leukocytes disintegrated and in some cases this happens immediately. Some patients suffering symptoms of the ear, nose and throat were found to also present clinical evidence for an allergic etiology involving food and based on the Bryans (1960) follow-up Cytotoxic testing of 107 of their patients this possibility was tested. They also presented a table that ranked the frequency that specific allergens were tested for and from which the percentage of positive or negative occurrences could be derived for their patient population of 107 (i.e., 122 tests for pork were 63.9% negative and 36.1% positive). Through diet management some form of relief for symptoms and syndromes studied in their population of 107 patients was achieved for 78.2%, no improvement for 12.8%, and for 9.0% it was unknown if any benefit was accomplished. The Bryans (1967, 1969, 1971, and 1972) continued to refine the description of their testing techniques and offered more detailed analyses of their findings from over 2500 patients tested.

The Cytotoxic Test Validated: Ulett M.D., Ph.D., (1973, 1974) while Director of the Missouri Institute of Psychiatry, a cooperative effort between the Missouri University Medical School and the Missouri Division of Mental Health, introduced a Cerebral Allergy program. This program included the use of Cytotoxic Testing as a diagnostic tool used to assist in the assessment of mental and psychological disorders. The Institute’s computer analyzed EEG’s and psychopharmacology programs had already received international recognition. The Bryans provided training and valuable consultation that expedited the establishment and early success for this program. The first studies followed the Cytotoxic Testing procedure of Black (1956) as described by the Bryans (1960, 1969, and 1971). Using the results of Cytotoxic Testing to design strict diet management programs Dr. Ulett built a highly successful program, which he continued until his retirement in the late 1990s, treating a broad range of clinical problems presented by patients seeking mental health services. Ulett and Perry (1974, 1975) made some significant observations while continuing to seek ways to improve and validate the Cytotoxic Testing procedure. Initial Cytotoxic Tests on subjects fasted since the previous evening were followed on a subsequent day with a second fasting Cytotoxic Test then a third challenge Cytotoxic Test at 1.5 - 2.0 hours after consuming foods known to be Cytotoxic Test positive based on the initial Cytotoxic Test results. Ulett and Perry (1974) reported that when subjects were challenged with cytotoxic (in vitro) positive tested foods, they showed a systemic (in vivo) leukocytosis that gave a direct correlation between the two different testing methods. That is, Cytotoxic Test positive food(s) when consumed following carefully managed procedures with proper controls caused the (in vivo) WBC to rise, and cytotoxic test negative food(s) caused no change in